New derivatives of testosterone



United States PatentO NEW DERIVATIVES OF TESTOSTERONE Poul Borrevang,Vanlose, Copenhagen, Denmark, as-

signor to Lovens Kemiske Fabrik Ved A. Kongsted, Ballerup, Denmark NoDrawing. Application November 17, 1959 Serial No. 853,446

Claims priority, application Great Britain November 19, 1958 6 Claims.(Cl. 260-3974) This invention relates to new and therapeutimlly usefulderivatives of testosterone, which derivativeshave the general formula:

l CE, I -42-6101;

in which R is selected from the group consisting of hydrogen, and thefollowing acyl groups: formyl, 'acetyl, propionyl, butyryl, andisobutyryl. Thus, the said derivatives aretestosterone-chloral-hemiacetal and certain esters thereof. It has beenfound that in aqueous suspensions these compounds are capable ofexerting androgenic and anabolic efiects which are substantially higherthan the same efiects of the propionate and the isobutyrate oftestosterone.

Testosterone-chloral-hemiacetal is very sparingly soluble in water, andin contact with water, it is decomposed into testosterone and chloral.For this reason aqueous suspensions of testosterone-chloral-hemiacetalare not suitable as saleable pharmaceutical preparations. When it isdesired to apply such suspensions for therapeutical purposes they shouldpreferably be prepared immediately be fore use. The esters of the saidhemiacetal are likewise very sparingly soluble in water, but are stablein contact with water. They show more protracted biological effects ofthe said kind than the esters of testosterone itself. Thus, for example,in aqueous suspensions the acetate of testosterone-chloral-hemiacetalshows more protracted andrw genie effect than testosterone isobutyrate.The initial, as well as the protracted, eflects of the acetate, thepropionate, and the isobutyrate of testosterone-chloral-hemiacetaldecrease in the sequence in which they are mentioned here.

The difierence in activity between testosterone-chloralhemiacetal andtestosterone propionate has been determined by tests according to themethod described by Eisenberg and Gordan in Journal of Pharmacology andExperimental Therapeutics, vol. 99 (1950), page 38. The test animals,castrated male rats, received daily, during 8' consecutive days, onesubcutaneous injection of an aqueous suspension containing varyingamounts of the new the other of the two compounds to be tested,,whereafter the weight of levator ani as well as of. prostata plusvesiculae seminalis were determined and' compared with the weight of thesame parts of control animals. results of the tests are specified in thefollowing table in The' 7 2,933,514 a ented Ap 1. 1 60 which the figuresin the columns A-E have the following meanings:

TABLE I A B o D E Androgem'c steroid compounds are preferablyadministered by intramuscular injections of aqueous suspensions of thecrystalline steroid compound. Thereby, protracted physiological efiectscan be obtained, and the degree of protraction depends upon the size ofthe crystals. In commercially available aqueous suspensions ofcrystalline testosterone isobutyrate a particle sizeof secures asuitable initial, maximum, and protracted effect.

It has been found that aqueous suspensions of the esters oftestosterone-chloral-hemiacetal, in which the particle size is about 10besides a satisfactory initial and maximum efiect, produce a moreprotracted efiect than that which can be obtained with aqueoussuspensionsof crystalline testosterone isobutyrate having a particlesize of 100 This superiority of the said esters has been ascer tained bycomparative tests applying the method described by Kupperman et al. inActa Endocrinogica, vol. 16 (1954), page 109. In this method thegrowth-promoting effect on the accessory sex glands of castrated malerats is used as a measure of the androgenic activity. The rats weredivided in groups of 10 animals, and each animal received a singleinjectioncontaining 2.0 mg. of testosterone isobutyrate ortestosterone-chloral-hemiacetal acetate, both substances in the form ofaqueous crystalline suspensions in which the particle size was 100 and1011., respectively. The animals were killed 4, 7, 14, and 21 days afterthe injection, and the weight of prostata plus vesiculae seminalis wasdetermined as the average weight of the glands from each group of 10animals. The results are specified I in Table H.

TABLE H Weight in mg. of prostate vesteulae seminalis By using aparticle size of only 10p. the suspension more easily passes through thecanula, and pains at the injection spot are diminished to such a degreethat it is possible toomitannaddition of a local anesthetic-to thesuspension.

Testosterone-chloral-hemiacetal can be produced by reacting testosteronewith chloral, preferably at room temperature as described in Example 1.The reaction can also be carried out at elevated temperatures, forexample at the boiling point of the reaction mixture. When the reactionis carried out at the boiling point, the chloral can be replaced bychloral hydrate. The esters of testosterone-chloral-hemiacetal withacids having two or more carbon atoms can be produced by reacting thesaid hemiacetal with the anhydride of the acid in question. The formiateof testosterone-chloral-hemiacetal can be produced by reacting thehemiacetal with the mixed anhydrideof formic acid and acetic acid. i

Example 1 Testosterone-chloral-hemiacetal.2.9 g. of testosterone weremixed at room temperature with a solution of 1.7 g. of anhydrous chloralin ml. of anhydrous benzene. By shfiing the mixture for a few minutes aclear solution was obtained, and after some further few minutes a solidprecipitate was formed. The precipitate was filtered off, washed withbenzene, and dried at room temperature. Thereby, 2.5 g. of a productwith melting point 194-196 C. were obtained, which was purified byrecrystallization from ethyl acetate. The recrystallizedtestosterone-chloralhemiacetal had M.P. 200-201" C. The ultravioletspectrum showed a maximum at 241. mp, e=l6,300, in ethanol. V t

1 'Calculated: C, 57.87%; H, 6.71%; Cl, 24.41%. Found: C, 57.62%; H,6.70%; CI, 24.61%.

' Example 2 benzoyl peroxide in 12 ml. of chloroform, and the mixturewas boiled with reflux for 2 hours and subsequently cooled to roomtemperature. Thereafter, the solution was washed 3 times with a diluteaqueous solution of sodium chloride, dried with Na SO and evaporated todryness in vacuo on a water bath. The crystalline residue was trituratedwith ether, and the solid material was filtered off and dried at roomtemperature. Thereby, 2.6 g. of testosterone-chloral-hemiacetal wereobtained with M.P. 192-194 C.

Example 5 perature for 16 hours and subsequently evaporated to dryphase.The latter was washed 3 times with water, whereby 300 mg. of a solidsubstance separated. The organic phase was dried with Na SO andthereafter evaporated to dryness in vacuo on a water bath. The drycrystalline residue was triturated with ethyl acetate, the mixture wasfiltered, and the filter cake dried at room temperature.

Thereby, 1.8 g. of testosterone-chloral-hemiacetal were obtained withM.P. 189-190 C.

Example 3 Testosterone-chloral-hemiacetal.A solution of 4.8 g. oftestosterone in 18 ml. of chloroform was mixed with a solution of 4.0 g.of chloral hydrate and 120 mg. of benzoyl peroxide in 13 ml. ofchloroform, and the solution was boiled with reflux for 2 hours andsubsequently cooled to room temperature. Thereafter, the solution waswashed 3 times with a dilute aqueous solution of sodium chloride, driedwith Na SQ;,'andvaporated to dryness in vacuoon a waterbath. -Thecrystalline residue was tritu rated with ether, and the solid materialwas filtered off and dried at room temperature. Thereby, 1.9 g. oftetosteronechloral-hemiacetal were obtained with M.P. 190-193" C.

'Example 4 I Testosterone-ehloral-hemiacetaL-A solution of 4.8 g. oftestosterone in 18 ml. of chloroform was mixed with a solution of 3.6 g.of anhydrouschloralandlSO mg- Of ness in vacuo on a water bath. Theresidue was triturated with ether, whereafter the solids were filteredofi, washed with ether, and dried; Thereby, 2.9 g. of the desiredacetate were obtained with M.P. l9'1-192 C. After recrystallization fromethyl acetate the M.P. was 192-193 C. The ultraviolet spectrum showed amaximum at 241 m e=16,400, in ethanol.

Caculated: C, 57.81%; H, 6.54%; Cl, 22.26%. Found: C, 57.69%; H, 6.61%;Cl, 22.46%.

Example 6 The propianate of testosterone-chloral-hemiacetal.3.0 g. oftestosterone-chloral-hemiacetal were dissolved in a mixture of 10 ml. ofpropionic acid anhydride and 10 ml. of pyridine. The solution was leftstanding at room temperature for 16 hours and was thereafter evaporatedin vacuo on a water bath to an oily materiaLwhich after a whilecrystallized. Petrol ether was added and the crystals were filtered offand dried. Thereby, 1.9 g. of the desired propionate were obtained withM.P. ISO-151 C. After recrystallization from methanol the M.P. was 151-152 C. The ultraviolet spectrum showed a maximum at 240 m e=l7,200, inethanol.

Calculated: C, 58.60%; H, 6.76%; C1, 21.63%.

7 Found: C, 58.55%; H, 6.74%; Cl, 21.59%.

Example 7 The isobutyrate of testosterone-chloral-hemiacetal.--3.0 g. oftestosterone-chloral-hemiacetal were dissolved in a mixture of 10 ml. ofisobutyric acid anhydride and 10 ml. of pyridine. The solution was leftstanding at room temperature for 16 hours, and was thereafter worked upas described in Example 6. Thereby, 1.8 g. of the desired isobutyratewere obtained with M.P. 126-127 C. After recrystallization from methanolthe M.P. was 127-128 C. The ultraviolet spectrum showed a maximum at 242mp, e='17,200, in ethanol.

Calculated: C, 59.35%; H, 6.97%; Cl, 21.03%. Found: ,C, 59.41%; H,7.21%; Cl, 21.03%.

Example 8 234 C-. (decomposition). After recrystallization from assignst e M411 raw-2 C-,

I claim: in which R is selected from the group consisting of hy- 1.Testosterone derivatives having the general formula: drogemformyl,acetyl, propionyl, butyryl, and isobutyryl.

CH; 2. Testosterone-chloral-hemiacetal.

5 3. The formate of testosterone-chloral-hemiacetal. CH: 0-0-0 01; 4.The acetate of testosterone-chloral-hemiacetal.

R 5. The propionate of testosterone-chlbral-hemiacetal.

6. The iso-butyrate of testosterone-chloral-hemiaceta1.

10 No references cited.

1. TESTOSTERONE DERIVATIVES HAVING THE GENERAL FORMULA: